DSMZ 1030 . SMITHELLA MEDIUM

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Culture medium recipe

Vol.
×
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.97 18.133
MgCl2 x 6 H2O 1.0 g 0.97 4.771
CaCl2 x 2 H2O 0.4 g 0.388 2.639
KH2PO4 0.4 g 0.388 2.851
Wolfe's mineral elixir 1.0 ml - -
Yeast extract 2.0 g 1.94 -
Trypticase peptone
(BD BBL)
2.0 g 1.94 -
Sodium resazurin
(0.1% w/v)
0.5 ml 4.8e-4 0.002
Na2CO3 1.0 g 0.97 9.151
Na-crotonate solution (1 M)
(1 M)
20.0 ml - -
Wolin's vitamin solution 10.0 ml - -
2-Mercaptoethanesulfonic acid
(coenzyme M)
0.5 g 0.485 3.41
Na2S x 9 H2O 0.3 g 0.291 1.212
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except carbonate, crotonate, coenzyme M, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into Hungate-type tubes or serum vials (e.g., 5 ml medium in Balch-type tubes) and autoclave. Add crotonate, coenzyme M, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of crotonate, coenzyme M and vitamins should be sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2.
2 Note: Use 10% (v/v) as inoculum.
Solution occurs in 23 other media (first defined in Medium 792).
Compound Amount Unit Conc. [g/L] Conc. [mM]
MgSO4 x 7 H2O 30.00 g 30 121.716
MnSO4 x H2O 5.00 g 5 29.583
NaCl 10.00 g 10 171.116
FeSO4 x 7 H2O 1.00 g 1 3.597
CoCl2 x 6 H2O 1.80 g 1.8 13.863
CaCl2 x 2 H2O 1.00 g 1 6.802
ZnSO4 x 7 H2O 1.80 g 1.8 6.26
CuSO4 x 5 H2O 0.10 g 0.1 0.401
AlK(SO4)2 x 12 H2O 0.18 g 0.18 0.379
H3BO3 0.10 g 0.1 1.617
Na2MoO4 x 2 H2O 0.10 g 0.1 0.413
(NH4)2Ni(SO4)2 x 6 H2O 2.80 g 2.8 7.089
Na2WO4 x 2 H2O 0.10 g 0.1 0.303
Na2SeO4 0.10 g 0.1 0.529
Distilled water 1000.00 ml - -
1 First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts.
Solution occurs in 3 other media (first defined in Medium 870).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Crotonic acid
(ALDRICH 113018)
0.86 g 86 998.955
NaOH
(10 N)
1.00 ml - -
Distilled water 9.00 ml - -
1 Dissolve crotonic acid in 8 ml distilled water, add around 1 ml of 10 N NaOH and adjust pH to around 7. Then add water to reach a volume of 10 ml. Sterilize by filtration under 100% N2 gas atmosphere.
Solution occurs in 357 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 25.03.22
Source: DSMZ
Medium type: Complex medium
Final pH: 7.0 - 7.2
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % N2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Yeast extract 1.94 -
Trypticase peptone 1.94 -
Crotonic acid 1.668 19.378
NH4Cl 0.97 18.133
Na2CO3 0.97 9.151
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Cultivation metadata from 0 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain